大鼠牙源性细胞复合nHAC/PLA支架的体外生物相容性研究

目的：探讨大鼠牙源性上皮细胞（rDECs）、牙源性间充质细胞（rDMCs）分层复合纳米羟基磷灰石-胶原/聚乳酸（nHAC/PLA）支架共生长的生物相容性.方法：①.应用酶消化法和差速消化法分离培养rDECs和rDMCs,经cytokeratin、vimentin免疫荧光染色鉴定其来源.②.应用扫描电镜观测rDECs、rDMCs分层复合nHAC/PLA支架后的生物相容性.结果：①.成功分离培养、鉴定大鼠牙源性上皮细胞和间充质细胞,免疫荧光染色鉴定rDECs、rDMCs分别表达cytokeratin、vimentin,证明rDECs、rDMCs分别来源于牙源性上皮组织、中胚层间充质.②.扫描电镜显示：rDECs、rDMCs复合nHAC/PLA共培养5d后,椭圆形的rDECs和长梭形的rDMCs胞体充分伸展,胞质中胶原纤维和基质分泌丰富,细胞与支架相互交织成网状.10d后,rDECs、rDMCs胞质中基质分泌更加丰富,覆盖部分细胞,并与支架相融合.结论：rDECs、rDMCs在nHAC/PLA支架上增殖、分化旺盛,具有良好的生物相容性.

The biocompatibility study after rat odontogenic cells were combined with nHAC/PLA scaffold in vitro

Objective：To investigate the biocompatibility after rat dental epithelial cells （rDECs） and rat dental mesenchymal cells （rDMCs） were combined with nHAC/PLA in mutualism.Method：①.Enzyme and differential digestions were performed to isolate and culture rDECs and rDMCs,and immunofluorescence staining against cytokeratin and vimentin were carried out to identify cell sources.②.Scanning electron microscope （SEM） was utilized to observe the biocompatibility after rDECs and rDMCs were combined with nHAC/PLA.Result：①.The rDECs and rDMCs were isolated,cultured and identified successfully.Immunofluorescence staining indicated cytokeratin and vimentin were expressed by rDECs and rDMCs respectively which demonstrated rDECs and rDMCs were from odontogenic epithelium and mesenchyme respectively.②.After co-cultured 5 d by rDECs ＋ rDMCs combined with nHAC/PLA,scanning electron microscope （SEM） indicated that elliptic rDECs and fusiform rDMCs extended thoroughly,collagen fibers and matrix in cytoplasms secreted abundantly,and cells interlace scaffolds to be reticulate.After co-cultured 10 d,matrix secretion in cytoplasms of rDECs and rDMCs was much more plentiful which covered partial cells and mixed together with scaffolds.Conclusion：The rDECs and rDMCs proliferated and differentiated productively on nHAC/PLA,which had favourable biocompatibility.